JP6248095B2 - 核酸の抽出及び貯蔵のための方法及び組成物 - Google Patents
核酸の抽出及び貯蔵のための方法及び組成物 Download PDFInfo
- Publication number
- JP6248095B2 JP6248095B2 JP2015510354A JP2015510354A JP6248095B2 JP 6248095 B2 JP6248095 B2 JP 6248095B2 JP 2015510354 A JP2015510354 A JP 2015510354A JP 2015510354 A JP2015510354 A JP 2015510354A JP 6248095 B2 JP6248095 B2 JP 6248095B2
- Authority
- JP
- Japan
- Prior art keywords
- solid matrix
- rna
- nucleic acid
- sample
- acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 150000007523 nucleic acids Chemical class 0.000 title claims description 44
- 108020004707 nucleic acids Proteins 0.000 title claims description 43
- 102000039446 nucleic acids Human genes 0.000 title claims description 43
- 238000000034 method Methods 0.000 title claims description 32
- 239000000203 mixture Substances 0.000 title claims description 31
- 238000003860 storage Methods 0.000 title claims description 27
- 238000000605 extraction Methods 0.000 title claims description 8
- 239000011159 matrix material Substances 0.000 claims description 53
- 239000007787 solid Substances 0.000 claims description 50
- 229920002678 cellulose Polymers 0.000 claims description 28
- 239000001913 cellulose Substances 0.000 claims description 28
- 239000003638 chemical reducing agent Substances 0.000 claims description 26
- 239000000523 sample Substances 0.000 claims description 23
- PZBFGYYEXUXCOF-UHFFFAOYSA-N TCEP Chemical compound OC(=O)CCP(CCC(O)=O)CCC(O)=O PZBFGYYEXUXCOF-UHFFFAOYSA-N 0.000 claims description 21
- 239000012472 biological sample Substances 0.000 claims description 17
- 239000000872 buffer Substances 0.000 claims description 17
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical group SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 claims description 15
- 102000004169 proteins and genes Human genes 0.000 claims description 15
- 108090000623 proteins and genes Proteins 0.000 claims description 15
- 210000004369 blood Anatomy 0.000 claims description 10
- 239000008280 blood Substances 0.000 claims description 10
- 210000004027 cell Anatomy 0.000 claims description 9
- 239000003398 denaturant Substances 0.000 claims description 9
- 239000003161 ribonuclease inhibitor Substances 0.000 claims description 9
- QIGBRXMKCJKVMJ-UHFFFAOYSA-N Hydroquinone Chemical compound OC1=CC=C(O)C=C1 QIGBRXMKCJKVMJ-UHFFFAOYSA-N 0.000 claims description 8
- 239000007983 Tris buffer Substances 0.000 claims description 8
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims description 8
- -1 vanadyl ribonucleoside Chemical compound 0.000 claims description 8
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 claims description 8
- NWVVVBRKAWDGAB-UHFFFAOYSA-N p-methoxyphenol Chemical group COC1=CC=C(O)C=C1 NWVVVBRKAWDGAB-UHFFFAOYSA-N 0.000 claims description 7
- 239000007987 MES buffer Substances 0.000 claims description 6
- 239000003795 chemical substances by application Substances 0.000 claims description 6
- 239000003112 inhibitor Substances 0.000 claims description 6
- 230000007774 longterm Effects 0.000 claims description 6
- 239000007993 MOPS buffer Substances 0.000 claims description 5
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 claims description 5
- 239000006172 buffering agent Substances 0.000 claims description 5
- ZJYYHGLJYGJLLN-UHFFFAOYSA-N guanidinium thiocyanate Chemical compound SC#N.NC(N)=N ZJYYHGLJYGJLLN-UHFFFAOYSA-N 0.000 claims description 5
- 210000001519 tissue Anatomy 0.000 claims description 5
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 claims description 4
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 4
- 210000003296 saliva Anatomy 0.000 claims description 4
- 210000002966 serum Anatomy 0.000 claims description 4
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 239000002342 ribonucleoside Substances 0.000 claims description 3
- SXGZJKUKBWWHRA-UHFFFAOYSA-N 2-(N-morpholiniumyl)ethanesulfonate Chemical compound [O-]S(=O)(=O)CC[NH+]1CCOCC1 SXGZJKUKBWWHRA-UHFFFAOYSA-N 0.000 claims description 2
- 239000004475 Arginine Substances 0.000 claims description 2
- 239000007995 HEPES buffer Substances 0.000 claims description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-O Htris Chemical compound OCC([NH3+])(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-O 0.000 claims description 2
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 2
- 235000010323 ascorbic acid Nutrition 0.000 claims description 2
- 229960005070 ascorbic acid Drugs 0.000 claims description 2
- 239000011668 ascorbic acid Substances 0.000 claims description 2
- 239000004202 carbamide Substances 0.000 claims description 2
- 229920002301 cellulose acetate Polymers 0.000 claims description 2
- 239000007979 citrate buffer Substances 0.000 claims description 2
- 239000003599 detergent Substances 0.000 claims description 2
- 239000003365 glass fiber Substances 0.000 claims description 2
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical group [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 claims description 2
- 125000003729 nucleotide group Chemical group 0.000 claims description 2
- 239000008363 phosphate buffer Substances 0.000 claims description 2
- DVLFYONBTKHTER-UHFFFAOYSA-N 3-(N-morpholino)propanesulfonic acid Chemical compound OS(=O)(=O)CCCN1CCOCC1 DVLFYONBTKHTER-UHFFFAOYSA-N 0.000 claims 3
- CNHDIAIOKMXOLK-UHFFFAOYSA-N toluquinol Chemical compound CC1=CC(O)=CC=C1O CNHDIAIOKMXOLK-UHFFFAOYSA-N 0.000 claims 3
- 238000002360 preparation method Methods 0.000 claims 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 56
- 108020004463 18S ribosomal RNA Proteins 0.000 description 25
- 108020004414 DNA Proteins 0.000 description 19
- 238000004458 analytical method Methods 0.000 description 12
- 230000000694 effects Effects 0.000 description 11
- 108091092328 cellular RNA Proteins 0.000 description 10
- KIUMMUBSPKGMOY-UHFFFAOYSA-N 3,3'-Dithiobis(6-nitrobenzoic acid) Chemical compound C1=C([N+]([O-])=O)C(C(=O)O)=CC(SSC=2C=C(C(=CC=2)[N+]([O-])=O)C(O)=O)=C1 KIUMMUBSPKGMOY-UHFFFAOYSA-N 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 7
- 102000006382 Ribonucleases Human genes 0.000 description 6
- 108010083644 Ribonucleases Proteins 0.000 description 6
- 239000011543 agarose gel Substances 0.000 description 6
- 108020004999 messenger RNA Proteins 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 229940123457 Free radical scavenger Drugs 0.000 description 5
- 239000000499 gel Substances 0.000 description 5
- 125000005842 heteroatom Chemical group 0.000 description 5
- 239000002516 radical scavenger Substances 0.000 description 5
- 108020004418 ribosomal RNA Proteins 0.000 description 5
- ZRALSGWEFCBTJO-UHFFFAOYSA-N Guanidine Chemical compound NC(N)=N ZRALSGWEFCBTJO-UHFFFAOYSA-N 0.000 description 4
- 150000001450 anions Chemical group 0.000 description 4
- 239000002738 chelating agent Substances 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 150000002430 hydrocarbons Chemical group 0.000 description 4
- 230000003278 mimic effect Effects 0.000 description 4
- 238000004321 preservation Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 238000002123 RNA extraction Methods 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000001962 electrophoresis Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 230000002349 favourable effect Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 230000006641 stabilisation Effects 0.000 description 3
- 238000011105 stabilization Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 108091033380 Coding strand Proteins 0.000 description 2
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- CHJJGSNFBQVOTG-UHFFFAOYSA-N N-methyl-guanidine Natural products CNC(N)=N CHJJGSNFBQVOTG-UHFFFAOYSA-N 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 102000035195 Peptidases Human genes 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 230000021839 RNA stabilization Effects 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 230000006750 UV protection Effects 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 150000008064 anhydrides Chemical class 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000013626 chemical specie Substances 0.000 description 2
- SWSQBOPZIKWTGO-UHFFFAOYSA-N dimethylaminoamidine Natural products CN(C)C(N)=N SWSQBOPZIKWTGO-UHFFFAOYSA-N 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 125000000524 functional group Chemical group 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 230000004224 protection Effects 0.000 description 2
- 238000004451 qualitative analysis Methods 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 238000003757 reverse transcription PCR Methods 0.000 description 2
- 239000008247 solid mixture Substances 0.000 description 2
- ZMZDMBWJUHKJPS-UHFFFAOYSA-N thiocyanic acid Chemical compound SC#N ZMZDMBWJUHKJPS-UHFFFAOYSA-N 0.000 description 2
- QVCQYYYTMIZOGK-UHFFFAOYSA-N (-)-butrin Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C(C(=O)CC(O2)C=3C=C(OC4C(C(O)C(O)C(CO)O4)O)C(O)=CC=3)C2=C1 QVCQYYYTMIZOGK-UHFFFAOYSA-N 0.000 description 1
- 125000003903 2-propenyl group Chemical group [H]C([*])([H])C([H])=C([H])[H] 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-M Acrylate Chemical compound [O-]C(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-M 0.000 description 1
- 108091029845 Aminoallyl nucleotide Proteins 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-M Bicarbonate Chemical compound OC([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-M 0.000 description 1
- ZOXJGFHDIHLPTG-UHFFFAOYSA-N Boron Chemical compound [B] ZOXJGFHDIHLPTG-UHFFFAOYSA-N 0.000 description 1
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 1
- LPADUVUWBIBZMU-UHFFFAOYSA-N Butrin Natural products OCC1OC(Oc2ccc3C(=O)CC(Oc3c2)c4ccc(OC5OC(CO)C(O)C(O)C5O)c(O)c4)C(O)C(O)C1O LPADUVUWBIBZMU-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 102000005853 Clathrin Human genes 0.000 description 1
- 108010019874 Clathrin Proteins 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 108010067770 Endopeptidase K Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 239000004593 Epoxy Substances 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 102100021699 Eukaryotic translation initiation factor 3 subunit B Human genes 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 101000896557 Homo sapiens Eukaryotic translation initiation factor 3 subunit B Proteins 0.000 description 1
- 101000988834 Homo sapiens Hypoxanthine-guanine phosphoribosyltransferase Proteins 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- PEEHTFAAVSWFBL-UHFFFAOYSA-N Maleimide Chemical compound O=C1NC(=O)C=C1 PEEHTFAAVSWFBL-UHFFFAOYSA-N 0.000 description 1
- 101710163270 Nuclease Proteins 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-L Phosphate ion(2-) Chemical compound OP([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-L 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 239000013614 RNA sample Substances 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000001266 acyl halides Chemical class 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- 150000001345 alkine derivatives Chemical class 0.000 description 1
- 150000001350 alkyl halides Chemical class 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 150000001502 aryl halides Chemical class 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 150000001540 azides Chemical class 0.000 description 1
- 239000006177 biological buffer Substances 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 229910052796 boron Inorganic materials 0.000 description 1
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 1
- 229940074995 bromine Drugs 0.000 description 1
- 229910052794 bromium Inorganic materials 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- QVCQYYYTMIZOGK-VQBAZXIRSA-N butrin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=C(C(=O)C[C@H](O2)C=3C=C(O[C@H]4[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O4)O)C(O)=CC=3)C2=C1 QVCQYYYTMIZOGK-VQBAZXIRSA-N 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 230000003196 chaotropic effect Effects 0.000 description 1
- XTEGARKTQYYJKE-UHFFFAOYSA-N chloric acid Chemical compound OCl(=O)=O XTEGARKTQYYJKE-UHFFFAOYSA-N 0.000 description 1
- 229940005991 chloric acid Drugs 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- QBWCMBCROVPCKQ-UHFFFAOYSA-N chlorous acid Chemical compound OCl=O QBWCMBCROVPCKQ-UHFFFAOYSA-N 0.000 description 1
- 229940077239 chlorous acid Drugs 0.000 description 1
- 229930193282 clathrin Natural products 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 239000004643 cyanate ester Substances 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 239000002274 desiccant Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-M dihydrogenphosphate Chemical compound OP(O)([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-M 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 230000007515 enzymatic degradation Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 239000011888 foil Substances 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000012948 isocyanate Substances 0.000 description 1
- 150000002513 isocyanates Chemical class 0.000 description 1
- 150000002540 isothiocyanates Chemical class 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 150000002596 lactones Chemical class 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 238000002493 microarray Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 150000002923 oximes Chemical class 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 229960004838 phosphoric acid Drugs 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- KCXFHTAICRTXLI-UHFFFAOYSA-N propane-1-sulfonic acid Chemical compound CCCS(O)(=O)=O KCXFHTAICRTXLI-UHFFFAOYSA-N 0.000 description 1
- 239000011814 protection agent Substances 0.000 description 1
- 239000003223 protective agent Substances 0.000 description 1
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 230000001603 reducing effect Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 125000005504 styryl group Chemical group 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 229940032330 sulfuric acid Drugs 0.000 description 1
- DHCDFWKWKRSZHF-UHFFFAOYSA-N sulfurothioic S-acid Chemical compound OS(O)(=O)=S DHCDFWKWKRSZHF-UHFFFAOYSA-N 0.000 description 1
- 150000008053 sultones Chemical class 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 150000003573 thiols Chemical class 0.000 description 1
- 238000005809 transesterification reaction Methods 0.000 description 1
- 229910052723 transition metal Inorganic materials 0.000 description 1
- 150000003624 transition metals Chemical class 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 229910052724 xenon Inorganic materials 0.000 description 1
- FHNFHKCVQCLJFQ-UHFFFAOYSA-N xenon atom Chemical compound [Xe] FHNFHKCVQCLJFQ-UHFFFAOYSA-N 0.000 description 1
- 238000004383 yellowing Methods 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
- B01J20/24—Naturally occurring macromolecular compounds, e.g. humic acids or their derivatives
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Analytical Chemistry (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Crystallography & Structural Chemistry (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Immunology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Sampling And Sample Adjustment (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Description
培養ヒトリンパ球細胞株(すなわち、Jurkat細胞)を全細胞RNA源として利用した。細胞を、表示した試薬で含浸させた7−mmセルロースディスク上で乾燥し、室温で10日間デシケーターキャビネット内に貯蔵し、細胞内核酸を標準的なプロトコルに従って電気溶出した。簡潔にいうと、ディスクを15μLのヌクレアーゼを含まない水中2mg/mLプロテイナーゼKで再水和して余分なタンパク質を除去し、〜30min乾燥した。パンチを1%Tris−ホウ酸−EDTA(TBE)アガロースゲルの個々のウェルに入れ、ホルムアミドを含有する1X Gel Loading Buffer II(Ambion)に懸濁させた。細胞内核酸を110ボルトで1〜2時間電気泳動させ、RNAとDNAをSYBR Gold(Invitrogen)で後染色し、Typhoon Imager(GE Healthcare)を用いて検出した。全ての機器と表面は、セルロースからの電気溶出中及びその後の細胞RNAの完全性を保存するためにRNAZap(Ambion)で処理した。RNA 6000 Nano Ladder(Agilent Technologies)及び筋肉から精製したヒト全RNA(Origene)を含めて内部標準をアガロースゲルに含ませて、RNaseの汚染をモニターすると共に対照のrRNAバンドを同定した。
この実施例の主たる目的は、セルロース紙上のRNAの保存に対する、各々の単一の因子の効果、及び試験した因子(例えば、キレート化剤、緩衝剤、pH、タンパク質変性剤、還元剤、及びペプチドRNase阻害剤)の組合せの効果を評価することであった。この実施例の追加の局面は、還元剤(DTT)の存在が、タンパク質変性剤の効果を高めることが可能であるか否かを評価することであった。
RNAを貯蔵するための鍵となる重要な成分が実施例2で同定された後、DTT及びSDSの単独又は組合せのRNAを保存する能力に対する効果、並びにGITC/DTTの組合せにフリーラジカル捕捉剤及びキレート化剤を追加することの、実施例2で好都合なRNA安定化特性を性能に対する効果を調べるために実施例3を立案した。
RNAを貯蔵するための追加の重要な成分が実施例3で同定された後、より良好な安定性を有し臭いがずっと少ない代わりの還元剤(TCEP)をDTTの代わりに用いることができるかどうか調べるために実施例4を立案した。本実施例に導入されたもう1つの因子はバナジルリボヌクレオシド複合体(VRC)、すなわち小分子RNase阻害剤であった。これらの変更を比較し、rRNAを安定化する能力を評価した。
実施例5は、選択組成物の30日間室温で貯蔵後の長期の性能を評価するために立案された。Jurkat細胞を直接セルロース紙試料上に付け、空気−乾燥して典型的な末端使用者の用途を模倣した。先の実施例1に記載したプロトコルに従って、全細胞RNAを電気溶出により1%アガロースゲル中に回収し、既知の標準に基づいて28s:18s rRNA含量を分析した。セルロース試料を30日間室温でデシケーターキャビネット内に貯蔵した後分析した。各バーの上の数字は28s対18s rRNAの比の値に対応する。28s:18sの比の値>1は一般にインタクトなRNAを示す。実施例5の結果は図5に示した。
実施例6は、様々な緩衝条件で新鮮な全血を有する選択紙組成物の性能を評価するために立案された。およそ50μLのラット全血を被験体の尾静脈から採取し、表示した緩衝剤成分で調製されたRNA安定化用の紙上にスポットして付けた。カードを乾燥し、周囲温度であるが制御された湿度(〜20%相対湿度)で5〜22日間貯蔵した。RNAを7mmセンターパンチから溶解緩衝剤中に抽出し、当技術分野で公知のプロトコルに従ってシリカ−膜スピンカラムに通して精製した。精製及び溶出後、RNA Integrity Number(RIN)を、RNA 6000 Pico LabChipsを用いるAgilent 2100 Bioanalyzerで測定した。慣習により、RIN>5は良好であるが、RIN>6はRT−PCR又はマイクロアレイ適用のような定量的下流分析に最良である。実施例6の結果は図6に示す。
実施例7は、選択乾燥マトリクス中に存在するUV抑制剤及び遊離基捕捉剤によるmRNA保護を立証するために立案された。DNAを含まない全Jurkat RNA(1μg)を、表示した成分を含有するRNA紙上に二つ組で(in duplicate)でスポットした。各々のカードを分割し、一方の半分を暗所に35℃で20時間保ち、他の半分はQ−SUN Xe−1 Xenon試験チャンバー内で20時間(35℃、0.3W/cm2、340nm)処理して、太陽光の全スペクトルを複製した(全エネルギー21.7kJ/m2)。各々の試料から1.2mmのパンチを採り、直接逆転写酵素反応液中に滴下してcDNAライブラリーを作成し、次いでこれをqPCRによりHPRT1及びクラスリンmRNAに特異的なプライマーに対して探索した(probed)。UVに露光した試料に対するサイクル閾値(CT)を暗所に貯蔵した未処理のつがいの一方のCTから差し引いた。実施例7の結果は図7に示す。
Whatman(商標)の31ETFセルロース系紙をTris緩衝液(pH7.4)中のGITCの存在下での増大する濃度のTCEP又はDTTに浸した。セルロース系紙を湿度調節をしないで室温で貯蔵した。5、19、及び105日に、5,5’−ジチオ−ビス(2−ニトロ安息香酸)(「DTNB」)を各々の紙試料上に載せた。活性な還元剤の存在下で、瞬時の黄変が観察された。周囲条件下で105日までの貯蔵において、全ての濃度のTCEP溶液を塗布したセルロース紙は活性のままであり、紙の白色から黄色への色の変化で示されるようにDTNBを還元することができた。DTTに浸した紙試料はDTNBを還元することができなかったので、紙の色は白色のままであった。これらの点はその意味が書面で表わされていないので、本明細書には含めないが、審査官から要請があれば提出する用意がある。DTNBの還元に関連する化学反応はClineら(2004)Biochemistry 43:15195−15203に見られる。
31ETFセルロース紙試料はいろいろな濃度の還元剤TCEP又はDTTを含むTris緩衝液(pH7.4)中にGITCを含有していた。紙試料を、1)21℃、10%相対湿度、2)21℃、80%相対湿度、3)41℃、10%相対湿度といういろいろな条件で貯蔵した。
さらにいろいろな緩衝液(Tris、pH7.4、MES、pH6.2、及びMPOS、pH7.0)中にGITC及びMEHQを含むTCEP組成物、並びに緩衝液を含まない対照試料を調製した。次に、セルロース系紙を各々上記溶液の異なる一つを塗布し、通気した50℃のオーブンですばやく乾燥し、アルミニウムホイルバッグ内に乾燥剤と共に密封して湿気を低く維持し、次に4℃、室温、又は41℃で貯蔵した。
Claims (15)
- 試料からの核酸の抽出及び貯蔵のための固体マトリクスであって、1種以上のタンパク質変性剤、1種以上の還元剤、UV抑制剤及び緩衝剤を含む組成物が固体マトリクス中に含浸されて乾燥状態で存在しており、固体マトリクスが、セルロース、酢酸セルロース、ガラス繊維又はこれらの組合せを含む多孔性マトリクスであり、タンパク質変性剤が、グアニジニウム塩酸塩、チオシアン酸グアニジニウム(GITC)、アルギニン、ドデシル硫酸ナトリウム(SDS)、尿素及びこれらの組合せから選択され、還元剤が、ジチオスレイトール(DTT)、2−メルカプトエタノール(2−ME)、トリス(2−カルボキシエチル)ホスフィン(TCEP)及びこれらの組合せから選択され、UV抑制剤が、ヒドロキノンモノメチルエーテル(MEHQ)、ヒドロキノン(HQ)、トルヒドロキノン(THQ)、アスコルビン酸及びこれらの組合せから選択される、固体マトリクス。
- タンパク質変性剤がGITCであり、還元剤がTCEPであり、UV抑制剤がMEHQ、THQ又はこれらの組合せから選択される、請求項1記載の固体マトリクス。
- 固体マトリクス中に存在する組成物がさらにRNase阻害剤を含む、請求項1又は請求項2記載の固体マトリクス。
- 固体マトリクスが周囲条件下において乾燥形態での核酸の長期貯蔵を可能にする、請求項1乃至請求項3のいずれか1項記載の固体マトリクス。
- 核酸がRNA、DNA又はこれらの組合せである、請求項1乃至請求項4のいずれか1項記載の固体マトリクス。
- 核酸がRNAである、請求項5記載の固体マトリクス。
- 多孔性マトリクスがセルロース紙である、請求項1乃至請求項5のいずれか1項記載の固体マトリクス。
- 緩衝剤が、2−アミノ−2−ヒドロキシメチル−プロパン−1,3−ジオール(Tris)、2−(N−モルホリノ)エタンスルホン酸(MES)、3−(N−モルホリノ)プロパンスルホン酸(MOPS)、4−(2−ヒドロキシエチル)−1−ピペラジンエタンスルホン酸(HEPES)、クエン酸緩衝剤及びリン酸緩衝剤からなる群から選択される、請求項1乃至請求項7のいずれか1項記載の固体マトリクス。
- 緩衝剤が、3〜8のpHを有する、請求項8記載の固体マトリクス。
- RNase阻害剤がバナジルリボヌクレオシド複合体(VRC)、ヌクレオチド類似体又は市販のRNase阻害剤である、請求項3記載の固体マトリクス。
- 固体マトリクスが多孔性のセルロース性マトリクスであり、かつ
a)タンパク質変性剤がGITC、洗剤又はこれらの組合せであり、
b)還元剤がDTT、TCEP又はこれらの組合せであり、かつ
c)緩衝剤がTris、MES又はMOPSである、請求項1記載の固体マトリクス。 - 核酸を試料から抽出し貯蔵する方法であって、
a)請求項1乃至請求項11のいずれか1項記載の固体マトリクスを用意する工程と、
b)試料を固体マトリクスに適用して核酸を収集する工程と、
c)固体マトリクスを乾燥する工程と、
d)周囲条件下において核酸を乾燥状態で固体マトリクス上に貯蔵する工程と
を含む方法。 - 当該方法がさらに、固体マトリクスから核酸を回収することを含む、請求項12記載の方法。
- 生物学的試料が血液、血清、組織、唾液又は細胞である、請求項12記載の方法。
- 試料が精製した核酸試料又は組織培養細胞調製物である、請求項12記載の方法。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US13/460,076 | 2012-04-30 | ||
US13/460,076 US9044738B2 (en) | 2012-04-30 | 2012-04-30 | Methods and compositions for extraction and storage of nucleic acids |
PCT/US2013/038576 WO2013165870A1 (en) | 2012-04-30 | 2013-04-29 | Methods and compositions for extraction and storage of nucleic acids |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2017225326A Division JP6847365B2 (ja) | 2012-04-30 | 2017-11-24 | 核酸の抽出及び貯蔵のための方法及び組成物 |
Publications (3)
Publication Number | Publication Date |
---|---|
JP2015518569A JP2015518569A (ja) | 2015-07-02 |
JP2015518569A5 JP2015518569A5 (ja) | 2016-06-09 |
JP6248095B2 true JP6248095B2 (ja) | 2017-12-13 |
Family
ID=49477845
Family Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2015510354A Active JP6248095B2 (ja) | 2012-04-30 | 2013-04-29 | 核酸の抽出及び貯蔵のための方法及び組成物 |
JP2017225326A Active JP6847365B2 (ja) | 2012-04-30 | 2017-11-24 | 核酸の抽出及び貯蔵のための方法及び組成物 |
JP2020036905A Pending JP2020092709A (ja) | 2012-04-30 | 2020-03-04 | 核酸の抽出及び貯蔵のための方法及び組成物 |
Family Applications After (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2017225326A Active JP6847365B2 (ja) | 2012-04-30 | 2017-11-24 | 核酸の抽出及び貯蔵のための方法及び組成物 |
JP2020036905A Pending JP2020092709A (ja) | 2012-04-30 | 2020-03-04 | 核酸の抽出及び貯蔵のための方法及び組成物 |
Country Status (8)
Country | Link |
---|---|
US (1) | US9044738B2 (ja) |
EP (1) | EP2844755B1 (ja) |
JP (3) | JP6248095B2 (ja) |
CN (1) | CN104254776B (ja) |
CA (1) | CA2870038C (ja) |
ES (1) | ES2609763T3 (ja) |
IN (1) | IN2014DN08223A (ja) |
WO (1) | WO2013165870A1 (ja) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2016527897A (ja) * | 2013-08-16 | 2016-09-15 | ゼネラル・エレクトリック・カンパニイ | 核酸の抽出及び保存のための方法及び組成物 |
Families Citing this family (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9480966B2 (en) | 2012-04-30 | 2016-11-01 | General Electric Company | Substrates and methods for collection, stabilization and elution of biomolecules |
US10829291B2 (en) * | 2012-06-20 | 2020-11-10 | Thermo Fischer Scientific Baltics UAB | Method to prevent silica-based column aging |
CN106572650B (zh) | 2014-06-10 | 2021-08-31 | 生物马特里卡公司 | 在环境温度下稳定凝血细胞 |
CN107076764A (zh) * | 2014-11-20 | 2017-08-18 | 通用电气医疗集团英国有限公司 | 检测稳定在固体载体材料上的痴呆和阿尔茨海默病相关生物标记 |
CN108473932B (zh) | 2015-09-09 | 2022-07-15 | 集联健康有限公司 | 用于样品收集、稳定化和保存的系统、方法和装置 |
US10000742B2 (en) * | 2015-11-19 | 2018-06-19 | General Electric Company | Device and method of collection for RNA viruses |
EP4242628A3 (en) | 2015-12-08 | 2023-11-08 | Biomatrica, INC. | Reduction of erythrocyte sedimentation rate |
FI3430373T3 (fi) * | 2016-03-18 | 2023-08-18 | Hummingbird Diagnostics Gmbh | Rna-fraktioiden puhdistus käyttämällä hydrofilistä polymeeristä materiaalia |
CN105961375A (zh) * | 2016-07-13 | 2016-09-28 | 北京中科唯新生物医学研究所有限公司 | 一种唾液保存液及其制备方法和用途 |
CN110072989A (zh) * | 2016-10-24 | 2019-07-30 | 生物马特里卡公司 | 核酸在纸上的稳定化 |
CN210383905U (zh) | 2017-01-10 | 2020-04-24 | 集联健康有限公司 | 一种用于从受试者收集流体样品的装置以及运输套筒 |
CN109182332A (zh) * | 2018-09-30 | 2019-01-11 | 浙江天杭生物科技股份有限公司 | 一种从临床样品中快速简便提取核酸的方法 |
CA3172263A1 (en) * | 2020-05-14 | 2021-11-18 | Joerg Hucklenbroich | Isolation of nucleic acids at elevated temperatures |
WO2022045303A1 (ja) | 2020-08-28 | 2022-03-03 | 花王株式会社 | Rnaの保存方法 |
GB202211645D0 (en) | 2022-08-09 | 2022-09-21 | Hemodx As | Kit |
Family Cites Families (59)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4540506A (en) | 1983-04-15 | 1985-09-10 | Genex Corporation | Composition for cleaning drains clogged with deposits containing hair |
CA1255197A (en) * | 1984-09-24 | 1989-06-06 | Joseph L. Orlik | Leukocyte differentiation composition and method |
US5173422A (en) | 1986-08-22 | 1992-12-22 | Miles Inc. | Monoclonal antibodies specific for human glycoalbumin |
US5756126A (en) | 1991-05-29 | 1998-05-26 | Flinders Technologies Pty. Ltd. | Dry solid medium for storage and analysis of genetic material |
US5807527A (en) | 1991-05-29 | 1998-09-15 | Flinders Technologies Pty. Ltd. | Solid medium and method for DNA storage |
US6627226B2 (en) | 1988-10-05 | 2003-09-30 | Whatman, Inc. | Dry solid medium for storage and analysis of genetic material |
WO1996039813A1 (en) | 1995-06-07 | 1996-12-19 | Flinders Technologies Pty. Ltd. | Dry solid medium for storage and analysis of genetic material |
US5981218A (en) | 1995-08-09 | 1999-11-09 | Bristol-Myers Squibb Company | Isolated nucleic acid molecules useful as leukemia markers and in breast cancer prognosis and encoded polypeptides |
US5939259A (en) | 1997-04-09 | 1999-08-17 | Schleicher & Schuell, Inc. | Methods and devices for collecting and storing clinical samples for genetic analysis |
US6204375B1 (en) | 1998-07-31 | 2001-03-20 | Ambion, Inc. | Methods and reagents for preserving RNA in cell and tissue samples |
DE19836559A1 (de) | 1998-08-12 | 2000-03-23 | Antigen Gmbh | Gefäß zur Entnahme von Blut |
US20010039010A1 (en) | 1998-09-03 | 2001-11-08 | Leigh Alexander Burgoyne | Sample collection medium incorporating material for sample visualization |
WO2000075302A2 (en) | 1999-04-30 | 2000-12-14 | Cyclops Genome Sciences Limited | Isolation of nucleic acid |
AU4353500A (en) | 1999-04-30 | 2000-11-17 | Whatman, Inc. | Substrate including anionic detergent for purifying nucleic acid |
US6664379B1 (en) | 1999-09-24 | 2003-12-16 | Ambion, Inc. | Nuclease inhibitor cocktail |
DE10006662A1 (de) | 2000-02-15 | 2001-08-23 | Antigen Produktions Gmbh | Gefäß zur Nukleinsäureanalytik |
US6602718B1 (en) | 2000-11-08 | 2003-08-05 | Becton, Dickinson And Company | Method and device for collecting and stabilizing a biological sample |
US7531362B2 (en) | 2001-06-07 | 2009-05-12 | Medmira Inc. | Rapid diagnostic assay |
EP1423514A2 (en) | 2001-09-05 | 2004-06-02 | WHATMAN plc | Stable storage of proteins |
GB0129816D0 (en) * | 2001-12-13 | 2002-01-30 | The Technology Partnership Plc | Testing device for chemical or biochemical analysis |
US7482116B2 (en) | 2002-06-07 | 2009-01-27 | Dna Genotek Inc. | Compositions and methods for obtaining nucleic acids from sputum |
EP1552005A4 (en) | 2002-07-10 | 2010-12-15 | Massachusetts Inst Technology | APPARATUS AND METHOD FOR ISOLATING A NUCLEIC ACID FROM A SAMPLE |
DE10231659B4 (de) | 2002-07-12 | 2006-01-19 | Preanalytix Gmbh | Zusammensetzung zum Binden von Nukleinsäure an eine Festphase |
GB0217963D0 (en) | 2002-08-02 | 2002-09-11 | Cyclops Genome Sciences Ltd | Purification of nucleic acids |
US7267950B2 (en) * | 2003-05-01 | 2007-09-11 | Veridex, Lcc | Rapid extraction of RNA from cells and tissues |
EP1628906A1 (en) * | 2003-05-23 | 2006-03-01 | Gyros Patent Ab | Fluidic functions based on non-wettable surfaces |
US7250270B2 (en) | 2003-06-13 | 2007-07-31 | Ambion, Inc. | Methods and compositions for preparing tissue samples for RNA extraction |
US20050042656A1 (en) | 2003-07-09 | 2005-02-24 | Davis James C. | Room temperature elution of nucleic acids |
JP3714940B2 (ja) | 2003-11-07 | 2005-11-09 | 株式会社日立ハイテクノロジーズ | Rna抽出方法、および生体材料の分析方法 |
US20060099567A1 (en) | 2004-04-08 | 2006-05-11 | Biomatrica, Inc. | Integration of sample storage and sample management for life science |
US20080176209A1 (en) | 2004-04-08 | 2008-07-24 | Biomatrica, Inc. | Integration of sample storage and sample management for life science |
ATE497837T1 (de) | 2004-04-09 | 2011-02-15 | Vivebio Llc | Vorrichtungen und verfahren für abnahme, lagerung und transport von biologischen proben |
DE102004021780B4 (de) | 2004-04-30 | 2008-10-02 | Siemens Ag | Verfahren und Anordnung zur DNA-Isolierung mit Trockenreagenzien |
EP1758937B1 (en) * | 2004-05-24 | 2009-09-02 | Genvault Corporation | Stable protein storage and stable nucleic acid storage in recoverable form |
US7776530B2 (en) | 2004-06-29 | 2010-08-17 | Wallac Oy | Integrated nucleic acid analysis |
US20090208919A1 (en) | 2005-01-21 | 2009-08-20 | Argylla Technologies, Llp | Particle matrix for storage of biomolecules |
CN101175853A (zh) | 2005-03-16 | 2008-05-07 | Dna吉诺特克股份有限公司 | 用于保存来自体液的核酸的组合物和方法 |
GB0505909D0 (en) | 2005-03-23 | 2005-04-27 | Univ Leeds | Formulations |
US20070015165A1 (en) | 2005-07-13 | 2007-01-18 | Sigma-Aldrich Co. | Method for the isolation of RNA from biological sources |
JP4699868B2 (ja) * | 2005-11-04 | 2011-06-15 | 株式会社日立ハイテクノロジーズ | 核酸精製方法及び核酸精製器具 |
EP1963526A4 (en) | 2005-12-09 | 2009-11-18 | Promega Corp | PURIFICATION OF NUCLEIC ACID WITH A BINDING MATRIX |
US8163535B2 (en) | 2006-06-26 | 2012-04-24 | Blood Cell Storage, Inc. | Devices and processes for nucleic acid extraction |
US8080645B2 (en) * | 2007-10-01 | 2011-12-20 | Longhorn Vaccines & Diagnostics Llc | Biological specimen collection/transport compositions and methods |
EP2191897B1 (en) * | 2007-06-21 | 2014-02-26 | Gen-Probe Incorporated | Instrument and receptacles for performing processes |
US20090053704A1 (en) * | 2007-08-24 | 2009-02-26 | Natalia Novoradovskaya | Stabilization of nucleic acids on solid supports |
AU2008329833B2 (en) * | 2007-11-30 | 2014-04-17 | Global Life Sciences Solutions Usa Llc | Method for isolation of genomic DNA, RNA and proteins from a single sample |
US20100209957A1 (en) | 2008-06-20 | 2010-08-19 | Genvault Corporation | Biosample storage devices and methods of use thereof |
CN102177237B (zh) | 2008-09-12 | 2013-10-30 | 金沃特公司 | 用于贮存和稳定生物分子的基质和介质 |
US9415392B2 (en) * | 2009-03-24 | 2016-08-16 | The University Of Chicago | Slip chip device and methods |
US8519125B2 (en) * | 2009-05-11 | 2013-08-27 | Biomatrica, Inc. | Compositions and methods for biological sample storage |
WO2011131720A1 (en) | 2010-04-20 | 2011-10-27 | Octapharma Ag | New stabilizing agent for pharmaceutical proteins |
EP2388312A1 (en) | 2010-05-17 | 2011-11-23 | Curetis AG | Universally applicable lysis buffer and processing methods for the lysis of bodily samples |
US9376709B2 (en) | 2010-07-26 | 2016-06-28 | Biomatrica, Inc. | Compositions for stabilizing DNA and RNA in blood and other biological samples during shipping and storage at ambient temperatures |
WO2012075471A1 (en) | 2010-12-04 | 2012-06-07 | Hui-Yu Liu | Rna stability enhancer |
US20120152743A1 (en) | 2010-12-17 | 2012-06-21 | General Electric Company | Method for electroeluting genetic material from dried samples |
GB201103257D0 (en) | 2011-02-25 | 2011-04-13 | Ge Healthcare Uk Ltd | Paper support and method of recovering biological material therefrom |
GB2496122A (en) | 2011-10-31 | 2013-05-08 | Ge Healthcare Bio Sciences Ab | Biological sample preservation on paper |
CN103173432B (zh) * | 2011-12-22 | 2020-08-04 | 通用电气公司 | 分离核酸的方法及装置 |
US9040675B2 (en) | 2012-04-30 | 2015-05-26 | General Electric Company | Formulations for nucleic acid stabilization on solid substrates |
-
2012
- 2012-04-30 US US13/460,076 patent/US9044738B2/en active Active
-
2013
- 2013-04-29 JP JP2015510354A patent/JP6248095B2/ja active Active
- 2013-04-29 WO PCT/US2013/038576 patent/WO2013165870A1/en active Application Filing
- 2013-04-29 ES ES13784927.9T patent/ES2609763T3/es active Active
- 2013-04-29 CA CA2870038A patent/CA2870038C/en active Active
- 2013-04-29 CN CN201380022584.6A patent/CN104254776B/zh active Active
- 2013-04-29 IN IN8223DEN2014 patent/IN2014DN08223A/en unknown
- 2013-04-29 EP EP13784927.9A patent/EP2844755B1/en active Active
-
2017
- 2017-11-24 JP JP2017225326A patent/JP6847365B2/ja active Active
-
2020
- 2020-03-04 JP JP2020036905A patent/JP2020092709A/ja active Pending
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2016527897A (ja) * | 2013-08-16 | 2016-09-15 | ゼネラル・エレクトリック・カンパニイ | 核酸の抽出及び保存のための方法及び組成物 |
Also Published As
Publication number | Publication date |
---|---|
IN2014DN08223A (ja) | 2015-05-15 |
CN104254776A (zh) | 2014-12-31 |
ES2609763T3 (es) | 2017-04-24 |
EP2844755A1 (en) | 2015-03-11 |
US9044738B2 (en) | 2015-06-02 |
WO2013165870A1 (en) | 2013-11-07 |
EP2844755B1 (en) | 2016-11-16 |
CA2870038C (en) | 2021-08-03 |
US20130289265A1 (en) | 2013-10-31 |
EP2844755A4 (en) | 2015-03-11 |
JP2018068303A (ja) | 2018-05-10 |
CN104254776B (zh) | 2016-09-28 |
JP2020092709A (ja) | 2020-06-18 |
JP6847365B2 (ja) | 2021-03-24 |
CA2870038A1 (en) | 2013-11-07 |
JP2015518569A (ja) | 2015-07-02 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP6248095B2 (ja) | 核酸の抽出及び貯蔵のための方法及び組成物 | |
US9040675B2 (en) | Formulations for nucleic acid stabilization on solid substrates | |
US9040679B2 (en) | Methods and compositions for extraction and storage of nucleic acids | |
CA2921286C (en) | Methods and compositions for extraction and storage of nucleic acids | |
DK2539449T3 (en) | PROCEDURE FOR PARALLEL ISOLATION AND CLEANING RNA AND DNA | |
JP6544860B2 (ja) | 固体基材上での核酸安定化のための配合物 | |
EP2971109B1 (en) | Methods for one step nucleic acid amplification of non-eluted samples | |
US9534214B2 (en) | Substrates and associated methods for elution of nucleic acids | |
CN103173432B (zh) | 分离核酸的方法及装置 | |
Olsen et al. | A rapid preparation procedure for laser microdissection-mediated harvest of plant tissues for gene expression analysis | |
WO2015108597A2 (en) | Substrates and associated methods for elution of nucleic acids |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20160414 |
|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20160414 |
|
A977 | Report on retrieval |
Free format text: JAPANESE INTERMEDIATE CODE: A971007 Effective date: 20170119 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20170207 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20170428 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20170707 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20170804 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20171024 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20171120 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 6248095 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
S111 | Request for change of ownership or part of ownership |
Free format text: JAPANESE INTERMEDIATE CODE: R313113 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |